Nigel Appleton Member
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Posts by Nigel Appleton

    In general, the LAMP isothermal nucleic acid amplification tests work better and faster with multiple primer pairs; and I would be VERY suspicious of the specificity of such a reaction operating at low temperatures (and I'm not aware of any DNA polymerases that operate efficiently at such temperatures. LAMP generally uses the Bst polymerase at 60+ deg C. Then again, I don't know everything, and haven't been in a PCR or LAMP lab for some years now)

    The reasons for using higher temperatures are related to stringency of primer binding, and the optimum temperature for the polymerase used - but PRIMARILY for the binding stringency. DNA and oligonucleotides are the very devil for non-specific binding at low temperatures; and of course ANY binding of a primer to a DNA sequence will set off a reaction

    And yes, I have helped develop a number of LAMP tests, now used commercially. Each has its optimum conditions, and even small departures from these (Mg++ concentration, primer sequences and concentrations, dNTP concentrations, reaction temperature, polymerase concentration, you name it) are just begging for non-specific reactions. These things are ALWAYS a balancing act between sensitivity (number of DNA or RNA targets detectable) and specificity,

    At their best LAMP-based NAATs are very fast, sensitive, and specific; and need require no expensive readers or thermocyclers, They are also much less sensitive to many of the inhibitors found in clinical samples, and sometimes need no more nucleic acid extraction/purification than a simple boiling step to lyse infected cells and virus particles and release the virus DNA or RNA

    Complete reaction mixes can be supplied freeze-dried, needing only to be reconstituted with sample extract in appropriate buffer, and heated. Freeze-dried reaction mixes can be made by the million in beaded form. I know this because I was responsible for the formulation and validation of the first ever freeze-drying formulations for LAMP tests.

    A long time ago now, as these things go.

    They do not always wear masks. They sometimes did in the past. Now they all have masks. In the past, you might see one or two people in a train car wearing masks, mainly for their own comfort, as I explained above. Now, everyone wears one. I am sure there is a tremendous difference in the infection rate for colds and influenza between the situation where one or two people wear masks, and everyone wears one. Previous years are not comparable.

    I spent a few weeks in Japan last year. About 30% of people on subway trains wore masks. On city streets, about 15%. On intercity trains and rural buses, and in rural towns and villages, very few. In restaurants and shops, virtually none. On the hiking trails, none at all.

    So far as I was aware, I caught no viruses; despite taking many subway journeys at busy times when whatever the opposite is of social distancing (?social cramming?) is practiced:)

    That ratio is changing mainly because young people are getting sick in larger proportions. That is not good news. It is also changing because therapy has improved. That is good.

    It will be interesting to see what the incidence and duration of sequelae such as lung damage, cardiovascular damage and neurological damage will be in various age cohorts. Maybe not good, but interesting.

    Me, I'm just keeping a reasonable distance from other people, not going to crowded places, wearing a mask in enclosed spaces where there are people, and I will take any vaccine that's passed safety and efficacy tests.

    Because I'm fit, well, and 72 - and I intend to stay that way as long as possible. And I don't want (if I DO happen to catch the damned thing) to pass it to anyone else.

    https://respectfulinsolence.co…fends-hydroxychloroquine/


    Did not take long for this professor in the Newsweek article to be hammered by this "colleague". Could not find who the author of the hit piece was, as he appears to be anonymous ("by Orac")? He gives the appearance of understanding Epidemiology, but his insulting, unscientific tone turned me off, so I did not read all the way to the end.


    From what I did read, he did not address how safe the drug was. If it is safe (and it is), and front line responders think it may work, then it should be freely available on an outpatient basis. End of story, until a preponderance of evidence proves otherwise.

    "Orac" is David Gorski, a respected surgeon


    https://en.wikipedia.org/wiki/David_Gorski

    For the record, it seems to me that most countries harbour people who like to use people as slaves. (other than sex slavery, which appears to be universal). In the UK, it is often a subset of agricultural gangmasters (those who organise teams of often-immigrant field workers in planting and harvest seasons) who are guilty. Sometimes rich people prefer not to pay servants and keep them in slavery. These slaves are poor (obviously!) , illegal immigrants, whose masters and mistresses confiscate any documents they may have There is often physical and sexual abuse in these situations.

    This sort of slavery certainly happens in Britain, although I do not think it's very common. The reports I've seen often involve foreign abusers from places where the practice is more common.. Occasionally. homegrown slavers are found, sometimes in Traveller communities tending to live tangentially to the law, if not outside it altogether.

    And I see Trump has "declared war" on testing!

    https://www.rawstory.com/2020/…s-all-out-war-on-testing/

    These are sad, - no, tragic -times indeed for the USA

    Unluckily the use of statins is correlated with dementia. It's also correlated with diabetes II and many other organ damages. Even worse there is no significant long time correlation for any coronary infarct prevention from statins..

    Thabks in advance

    Statins only show a good effect in short duration studies that completely neglect the long time damages. Thus I try to prevent as many people as possible from taking statins as you must take them for the rest of your live (big pharma hopes..).


    But some folks still believe a white frock makes a good and fear is the method of these goods.


    Could you give links, please, to the evidence that statin use is correlated with dementia? I couldn't find any


    Thanks in advance


    Oh, and you might be interested in this; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5418248/

    Review by editors dates back centuries. I do not know when peer-review by people other than the editor began. Anonymous peer-review where the author does not know who wrote the comments began in the 1930s, according to an Einstein biography I read. He was nonplussed. He wrote to the editor, 'What is this? Who wrote all these remarks about my paper?'


    It might have been the mid-1920s. I don't recall. But anyway, it is a recent custom, more honored in the breach than the observance.

    It is common nowadays for submitting authors to be asked to nominate reviewers. I cannot imagine a system more prone to induce "log-rolling" - a positive feedback system for work of dubious quality.

    Long ago, I had papers (fairly trivial, more like technical notes) independently reviewed by reviewers appointed by the journals to which the works were submitted. There were criticisms, mostly constructive, and the results were better publications. But then, my/our results were always replicable in other labs

    IMO if someone comes to this place claiming to have results, then they should report the results. Why make the claim if no report is intended?


    Repeated claims of results without showing results is at the very least annoying; and if done by the same person under serial avatars raises strong suspicions of trolling

    Nigel Appleton . There will be publication, but we can't jump the gun and publish here. Sadly.


    Oh, that's perfectly understandable. It's just that patience is not my strongest suit, and things seem to be moving quite fast now that there are multiple centres using a common (more-or-less) basis for development. The more positive results are reported, the more attempted replications and variations there will be, and there will be an acceleration of progress.

    And I would hate it if the field were suddenly to be shrouded in the fog of commercial confidentiality; although I expect it will happen.

    Is there any way we can see some numbers or graphs? Will there be any publication?

    This is extraordinarily encouraging to the interested scientifically literate layman for for several reasons;


    * We now have two teams of scientists getting and publishing reproducible methods of obtaining sustained excess heat

    * Both are using roughly similar techniques, i.e. exploiting nano-featured combinations of Ni/Pd (or Ni/Cu) with deuterium or hydrogen

    * Because of current intense interest in catalysis, there is a large and rapidly-expanding knowledge-base in the design and manufacture of nanosized metal particles, wires, bimetallics and composites - hence plenty of possibilities for optimisation.

    * In the work reported above, the materials, equipment, and pressures used (other than the nanoparticles themselves) are relatively accessible and in some cases really cheap (Cu and H, instead of Pd and D2;, internal pressures of the order of half-atmospheric, rather than hard vacuum)


    I'm tempted to predict that this field is about to attract much more interest from "mainstream" science.


    (And I can't wait for one of these teams to throw natural caution to the winds and start turning up the input power! =O=O)

    I uploaded some details and graphs from the replication at the Hokkaido University of Science. See p. 6:


    https://www.lenr-canr.org/acrobat/MizunoTsupplement.pdf


    Note: In the message above, I described this as an "independent replication." After reading through the materials and notes, I would say it is "semi-independent." But it is getting more independent by the day! I gather they are learning how to do it themselves, and so are the people at Hokkai PEEM Co., Ltd. I guess this means you need hands-on assistance at first. Perhaps people trying to replicate this should visit Mizuno, or visit Saito as the Hokkaido U. of Science.

    Jed, is the rate of heat loss through the enclosure walls linear with respect to changes of temperature within the enclosure? Constant? Is it correct to assume 150W loss over a wide range of internal temperatures?

    Did not Mizuno have failures as well as successes? If he used the same mesh and Pd, that might point rather firmly to the method of loading the mesh with Pd.

    Perhaps secondarily to any precleaning/ oxidation-during-storage issues.


    (I'm assuming the same or identical reactor(s) used. If not, a certain degree more mootness must apply to the my comments above! It is VERY difficult with so many factors to keep strict replication, let alone to get round to varying just one at a time)